N-amidinosarcosine

Endpoint summary

Description of key information

Four reliable acute toxicity studies on Creatine monohydrate are available.

Two oral studies in rats, one dermal study in the rat and one intraperitoneal study in mice. All studies are indicating no acute effect up to 2000 mg/kg body weight. A LD50 exceeding 2000 mg/kg body weight could be established. The corresponding calculated LD50 of creatine is exceeding 1758.36 mg/kg body weight. As none of the animals died during the studies the LD0 of creatine is exceeding 1758.36 mg/kg body weight. Therefore the LD50 of Creatine is expected to be above 2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997-8-27 to 1997-9-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: approx. 6 weeks old
- Weight at study initiation: female: average 145 g; male: average 180 g
- Fasting period before study: overnight prior to dosing until approximatetly 3-4 hours after administration of the test substance
- Housing: 3 animals per sex per cage
- Diet (e.g. ad libitum): free access
- Water (e.g. ad libitum): free access
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distilled

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 1 g test substance per 9 ml of water
- Justification for choice of vehicle: The vehicle was selected based on information provided by the sponsor

DOSAGE PREPARATION (if unusual): substance was ground into fine powder and 1 g was filled into the tube of a syringe; 9 ml water was drawn in; the syringe was shaken thoroughly to prepare the suspension; the suspension was administered immediately after preparation

Doses:

2000 mg/kg (20ml/kg) body weight

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: observations of clinical signs once daily; body weight: day 1 (pre-administration, 8 and 15)
- Necropsy of survivors performed: yes

Statistics:

no statistical analysis was performed

Preliminary study:

n.a.

Key result

Sex:

male/female

Dose descriptor:

LD0

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Remarks:

equals 1758.36 mg/kg bw Creatine (anhydrous)

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks:

equals 1758.36 mg/kg bw Creatine (anhydrous)

Mortality:

No mortality occurred

Clinical signs:

other: No clinical signs were detected

Gross pathology:

No abnormalities were found

Other findings:

none

Table 1: Body weights (gram)

Group/Sex	Animal	Day 1	Day 8	Day 15
Group 1/ Males (2000 mg/kg)	1	184	245	284
	2	188	243	292
	3	169	218	252
	Mean	180	235	276
	Standard deviation	10	15	21
	n	3	3	3
Group 1/ Females (2000 mg/kg)	1	147	177	203
	2	144	178	194
	3	144	192	223
	Mean	145	182	207
	Standard deviation	2	8	15
	n	3	3	3

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 value of creatine monohydrate in Wistar rats was established as exceeding 2000 mg/kg body weight. The corresponding LD50 of creatine is > 1758.36 mg/kg body weight.

Executive summary:

To assess the oral toxicity of creatine monohydrate a limit test based on the OECD guideline No. 423 "Acute oral toxicity-Acute toxic class method" was carried out. Creatine monohydrate in distilled water (1 g per 9 ml) was administered by oral gavage to three approx. 6 week old, fasted Wistar rats of each sex at a dose of 2000 mg/kg body weight. The animals were observed for 15 days. Macroscopic examination was performed after terminal sacrifice (day 15).

No mortality occurred and no clinical signs were noted. Body weight gain was considered as normal. No abnormalities were found at the macroscopic post mortem examination. The oral LD50 value of creatine monohydrate in Wistar rats was established as exceeding 2000 mg/kg body weight.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2004-10-5 to 2004-10-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Remarks:

IGS BR

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, D-97633 Sulzfeld
- Age at study initiation: approximately 8 weeks
- Weight at study initiation (mean): group 1 (dose 300 mg/kg b.w.): 175.3 g; group 2 (dose 300 mg/kg b.w.): 179.0 g; group 3 (dose 2000 mg/kg b.w.): 187.0 g; group 4 (dose 2000 mg/kg b.w.) 197.0 g
- Fasting period before study: evening before the administration and about 3 hours afterwards
- Housing: single caging
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): average: 22.1 °C
- Humidity (%): average 55.9 %
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

deionised

Details on oral exposure:

VEHICLE
- Concentration in vehicle: dependent on body weight; administration of 10 ml per kg b.w.; doses: 300 mg/kg and 2000 mg/kg b.w.
- Amount of vehicle (if gavage): 10 ml per kg b.w.
- Justification for choice of vehicle: the test substance could be disolved in water and water shall be used preferably, according to the guideline

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: as no prior information on the toxicity of the test substance was available, a starting dose of 300 mg/kg b.w. was chosen. The further proceeding was in accordance with the guideline.

Doses:

Starting dose: 300 mg/kg body weight
Second dose: 2000 mg/kg body weight

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical observations were performed within the periods 0-0.5, 0.5-1, 1-2, 2-4, 4-6 hours after administration and then at least once a day for a total of 2 weeks; body weights were determined before administration, 7 days p.a. and 14 days p.a.
- Necropsy of survivors performed: yes

Statistics:

no statistical analysis was performed

Preliminary study:

none

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks:

equal to 1758.36 mg/kg bw Creatine anhydride

Mortality:

All animals survived until the scheduled termination of the study

Clinical signs:

other: All animals were normal during the entire observation period

Gross pathology:

All animals were normal at the necroscopy 14 days after administration

Other findings:

no other findings

Table 1: Synopsis of the results

Dose (mg/kg)	Step No.	Animal Nos.	Exposed animals	Affected animals	Deceased animals
300	1	111, 112, 113	3	0	0
300	2	114, 115, 116	3	0	0
2000	3	121, 122, 123	3	0	0
2000	4	124, 125, 126	3	0	0

Table 2: Body weight and body weight gain

Dose mg/kg (Step No.)	Animal No.	Body weight				Body weight gain
		Before administration	7 days p.a.	14 days p.a.	death	0-7 days p.a.	7-14 days p.a.
300 (1)	111	170	203	231	-	33	28
	112	174	204	225	-	30	21
	113	182	217	239	-	35	22
	Mean	175.3	208.0	231.7	-	32.7	23.7
	SD	6.1	7.8	7.0	-	2.5	3.8
300 (2)	114	181	213	229	-	32	16
	115	177	214	246	-	37	32
	116	179	205	228	-	26	23
	Mean	179.0	210.7	234.3	-	31.7	23.7
	SD	2.0	4.9	10.1	-	5.5	8.0
2000 (3)	121	180	205	210	-	25	5
	122	184	212	227	-	28	15
	123	197	227	239	-	30	12
	Mean	187.0	214.7	225.3	-	27.7	10.7
	SD	8.9	11.2	14.6	-	2.5	5.1
2000 (4)	124	197	222	249	-	25	27
	125	198	224	236	-	26	12
	126	196	227	244	-	31	17
	Mean	197.0	224.3	243.0	-	27.3	18.7
	SD	1.0	2.5	6.6	-	3.2	7.6

Observations in life: normal at any time

Necroscopy findings: normal

Interpretation of results:

GHS criteria not met

Conclusions:

No toxic effects of the test substance were noted by signs in life and post mortem. No mortality occurred.

Executive summary:

To assess the oral toxicity of creatine monohydrate a limit test based on the OECD guideline No. 423 "Acute oral toxicity-Acute toxic class method" was conducted. Creatine monohydrate was administered once as a suspension in deionized water by oral gavage to groups of three fasted, approx. 8 weeks old animals (female Crl:CD(SD)IGS BR rats). A starting dose of 300 mg per kg body weight and a second dose of 2000 mg/kg body weight were given. The animals were observed for 14 days. No toxic effects of the test substance were noted. No mortality occurred. The oral LD50 value of creatine monohydrate in rats was established as exceeding 2000 mg/kg per body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-09-13 to 2011-09-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, D-97633 Sulzfeld.
- Age at study initiation: Approximately 8 weeks (males) and 12 weeks (females) at the time of administration.
- Weight at study initiation: males (mean): 282.2 g; females (mean): 230.4 g
- Fasting period before study: not reported
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 7 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Mean of 20.92 °C
- Humidity (%): Mean of 56.72 %
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 10 % of the estimated body surface
- % coverage: 10
- Type of wrap if used: cellulose patch (Pehazell, Hartmann AG), non irritating tape (Blenderm Wundpflaster, 3M), semi-occlusive dressing (Fixomull Stretch, Fa. Beiersdorf)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none; Residual test substance was wiped off using wet cellulose tissue, if necessary.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The amounts of the test substance were calculated and weighed for each individual using the body weight determined on the day of the administration.
- For solids, paste formed: no

Duration of exposure:

24 h

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: before administration, 7 days p.a., 14 days p.a.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, body weight gain; changes in skin, fur, eyes, the occurrence of secretions and excretions, autonomic activity, changes in gait, posture and the presence of convulsions

Statistics:

none

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks:

equals to 1758.36 mg/kg body weigh Creatine (anhydride)

Mortality:

All animals survived until the scheduled termination of the study.

Clinical signs:

other: General findings: All animals did not show any clinical signs during the entire observation period. Observations of skin condition: Exposed skin was not found to be altered by the test substance.

Gross pathology:

No abnormal findings were made in the animals at terminal necropsy.

Table 1: Body weights and body weight gain.

Individual data, mean and standard deviation SD.

Dose sex	Animal No.	Body weight (g)				Body weight gain (g)
2000 mg/kg male		before administr	7 days p.a.	14 days p.a.	death	0-7 days p.a.	7-14 days p.a.
	121	283	323	366	-	40	43
	122	295	335	378	-	40	43
	123	271	300	348	-	29	48
	124	277	296	334	-	19	38
	125	285	318	352	-	33	34
	Mean	282.2	314.4	355.6	-	32.2	41.2
	SD	9.0	16.3	16.9	-	8.8	5.4
2000 mg/kg female	126	226	251	269	-	25	18
	127	240	245	258	-	5	13
	128	232	233	240	-	1	7
	129	226	230	239	-	4	9
	130	228	243	252	-	15	9
	Mean	230.4	240.4	251.6	-	10.0	11.2
	SD	5.9	8.7	12.6	-	9.9	4.4

Interpretation of results:

GHS criteria not met

Conclusions:

No local or systemic toxic effects related to administration of the test substance were noted from clinical observations or post-mortem-examination at a dose of 2000 mg of the test substance per kg body weight. No mortality occurred.

Executive summary:

In an acute dermal toxicity study according the OECD guideline 402 groups of young adult rats (Crl:CD(SD)) (5/sex) were dermally exposed to Creatine Monohydrate (>=99 %) for 24 hours to 10 % of the body surface area at doses of 2000 mg/kg bw (limit test). Animals then were observed for 14 days. No local or systemic test substance related effects were noted from clinical observations or post-mortem examination at a dose of 2000 mg test substance per kg body weight. No mortality occurred. A dermal LD50 (males/females) > 2000 mg/kg body weight was determined. From the results of this study no classification is derived for Creatine Monohydrate according to the Directive 2001/59/EC.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

The acute oral toxicity of creatine monohydrate was assessed in two limit tests based on the OECD guideline No. 423 "Acute oral toxicity-Acute toxic class method" . No mortality occurred and no clinical signs were noted. The oral LD50 value of creatine monohydrate in Wistar rats was established as exceeding 2000 mg/kg body weight.

The acute dermal toxicity of Creatine Monohydrate was assessed in a limit test according the OECD guideline 402. No local or systemic test substance related effects were noted from clinical observations or post-mortem examination at a dose of 2000 mg test substance per kg body weight. A dermal LD50 (males/females) > 2000 mg/kg body weight was determined.

The intraperitoneal toxicity of Creatine Monohydrate was assessed in a limit test based on the OECD guideline 423 in mice (strain CD-1) except the route of dosing which was intraperitoneal instead of oral. No mortality occurred and no clinical signs were observed. The intraperitoneal LD50 value of creatine monohydrate in mice was established as exceeding 2000 mg/kg body weight.

As none of the animals died the LD0 is exceeding 2000 mg/kg body weight.

Toxicity by inhalation was not assessed as in case of the test substance Creatine the oral and dermal route are considered as likely routes of human exposure for workers and consumers. Besides that, Creatine has a very low vapor pressure and a high melting point, so the potential for the generation of inhalable forms is low. The particle size of Creatine is considered to be 114.02 µm. Therefore the particles are not considered to be inhalable (cut off size > 100 µm) and no acute inhalation test was performed.

Justification for classification or non-classification

No mortalities or clinical signs were observed at appropriate dose levels.

According Regulation (EC) No 1272/2008 LD50-doses of more than 2000 mg/kg are not considered to indicate oral or dermal acute toxicity.